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Microchimica Acta

, 186:133 | Cite as

Fluorometric determination of microRNA-122 by using ExoIII-aided recycling amplification and polythymine induced formation of copper nanoparticles

  • Yafang Tang
  • Mingxiu Liu
  • Zilin Zhao
  • Qing Li
  • Xuehua Liang
  • Jianniao TianEmail author
  • Shulin Zhao
Original Paper
  • 15 Downloads

Abstract

The authors describe a method for the determination of microRNA-122 by using terminal deoxynucleotidyl transferase (TdT). It is based on the use of polythymine and exonuclease III-aided cycling amplification. A 3′-phosphorylated hairpin probe 1 (H1) and a hairpin probe 2 (H2) were designed. In the presence of the microRNA, hybridization and enzymatic cleavage will occur and produce lots of 3′-hydroxylated ssDNA which can be tailed by TdT and converted into long polythymine (polyT) sequences. These can be used to synthesize copper nanoparticles (CuNPs) with fluorescence excitation/emission maxima at 350 nm/630 nm. This method shows good selectivity and high sensitivity with a linear response in the 1.00 × 102 fM and 1.00 × 106 fM microRNA concentration range and a 44 fM limit of detection. It was successfully applied to determination of microRNA in spiked serum samples.

Graphical abstract

A label-free and highly sensitive fluorometric method is described for the assay of microRNA on the basis of target-triggered two-cycle amplification and combining with terminal TdT. It produces a series superlong polyT that can be used for synthesis of copper nanoclusters (CuNCs) displaying red fluorecence.

Keywords

MicroRNA PolyT-CuNPs Polymerization Target recycling 

Notes

Compliance with ethical standards

The authors got the permission for using serum sample of human volunteers from Guilin’s Fifth People’s Hospital (China) according to institutional guidelines. All animal procedures and all experiments were performed in accordance with the Guidelines for Care and Use of Laboratory Animals of Guangxi Normal University (Guilin, China) and approved by the Animal Ethics Committee of China.

Supplementary material

604_2019_3237_MOESM1_ESM.docx (2.2 mb)
ESM 1 (DOCX 2.15 mb)

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Copyright information

© Springer-Verlag GmbH Austria, part of Springer Nature 2019

Authors and Affiliations

  • Yafang Tang
    • 1
  • Mingxiu Liu
    • 1
  • Zilin Zhao
    • 1
  • Qing Li
    • 1
  • Xuehua Liang
    • 1
  • Jianniao Tian
    • 1
    Email author
  • Shulin Zhao
    • 1
  1. 1.Key Laboratory for the Chemistry and Molecular Engineering of Medicinal Resources (Ministry of Education of China)School of Chemistry and Pharmaceutical Science of Guangxi Normal UniversityGuilinChina

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